Quantifying the reproduction of Bacillus thuringiensis HD1 in cadavers and live larvae of Plutella xylostella.

The Bacillus cereus group comprises a range of micro-organisms with diverse habits, including gut commensals, opportunistic pathogens and soil saprophytes. Using quantitative microbiological methods we tested whether Bacillus thuringiensis (Bt) could reproduce in cadavers of Plutella xylostella killed by Bt, or in the gut of live insects, or be transmitted vertically from females to their offspring. We also tested whether diverse Bt strains could grow in high nutrient broth at a pH similar to that in the larval midgut. Low levels of reproduction were found in insect cadavers but there was no evidence of vertical transmission, or of significant reproduction in live insects. Four strains of B. thuringiensis var. kurstaki and one of B. thuringiensis var. tenebrionis were found to be capable of growth at high pH. Greater spore recovery rates in frass were found in hosts that were resistant or tolerant of infection. We concluded that that spores recovered in frass represent, in general, an ungerminated fraction of ingested inoculum and that germination rates are reduced in unsuitable hosts.

Social and sexual behaviours aid transmission of bacteria in birds.

Understanding the behavioural mechanisms that mediate pathogen transmission in social hosts like birds could provide the empirical bases for explaining the epidemiological dynamics of zoonotic infections in vertebrates. By experimentally infecting the feathers and cloaca of captive zebra finches (Taeniopygia guttata), with the bacterium Bacillus licheniformis PWD1 (BL), we examined the self-contamination and horizontal transmission of birds sharing the same environment. We also examined whether sexual transmission of bacteria is gender biased. Our results show that bacteria placed on the plumage of the birds lead to self and allo-infections of the bird guts, possibly through preening behaviours and bacterial ingestion. Furthermore, we found that sexual transmission of the bacteria was asymmetrical, being higher when males are the transmitting sex. Our results suggest the existence of an oral-faecal-genital route of bacterial transmission for avian hosts, wherein bacteria present on feathers infect their host guts through self and allo-preening and bacterial ingestion. Gut bacteria can then be transmitted sexually with transmission rate being higher when males are the infected sex.

Tonometers and infectious risk: myth or reality? Efficacy of different disinfection regimens on tonometer tips.

PURPOSE: To evaluate the adequacy of common disinfection regimens for disposable tonometer tips and assess if disinfection of reusable prisms or the use of disposable tips is preferable. METHODS: We used disposable tonometer tips, using the same material and tip diameter of standard Goldmann tonometer prism. Strains of Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilisand Candida albicanswere tested according to the European standard guidelines for disinfectants test. Antimicrobial effectiveness of the following disinfection practices has been assessed: dry wipe, Minuten wipes (Alpro), soaking in 3% hydrogen peroxide, 0.5% benzalkonium chloride, and 0.5% Pantasept for 1, 5, and 15 min. All tests have been performed three times and all conditions tested in duplicate. RESULTS: Dry wiping and 1 min soak in 3% hydrogen peroxide were ineffective on all microrganisms. Minuten wipes, 1 min soak in 0.5% benzalkonium chloride or 3% hydrogen peroxide were ineffective on B. subtilis. 0.5% Pantasept soak was effective in 1 min for all microrganisms tested, whereas 3% hydrogen peroxide and 0.5% benzalkonium chloride soaks were effective when performed for at least 5 min. B. subtiliswas the most resistant organism to disinfectant regimes at 1 min time. CONCLUSIONS: Results of our study demonstrate a relative disinfection efficacy for the different evaluated regimens, provided that correct exposure times are adopted for the chosen disinfectants, a condition difficult to ensure in a busy clinic setting. We conclude that disposable prism tonometry provides a safe alternative to Goldmann tonometry.

Nosocomial pseudoepidemic caused by Bacillus cereus traced to contaminated ethyl alcohol from a liquor factory.

From September 1990 to October 1990, 15 patients who were admitted to four different departments of the National Taiwan University Hospital, including nine patients in the emergency department, three in the hematology/oncology ward, two in the surgical intensive care unit, and one in a pediatric ward, were found to have positive blood (14 patients) or pleural effusion (1 patient) cultures for Bacillus cereus. After extensive surveillance cultures, 19 additional isolates of B. cereus were recovered from 70% ethyl alcohol that had been used as a skin disinfectant (14 isolates from different locations in the hospital) and from 95% ethyl alcohol (5 isolates from five alcohol tanks in the pharmacy department), and 10 isolates were recovered from 95% ethyl alcohol from the factory which supplied the alcohol to the hospital. In addition to these 44 isolates of B. cereus, 12 epidemiologically unrelated B. cereus isolates, one Bacillus sphaericus isolate from a blood specimen from a patient seen in May 1990, and two B. sphaericus isolates from 95% alcohol in the liquor factory were also studied for their microbiological relatedness. Among these isolates, antibiotypes were determined by using the disk diffusion method and the E test, biotypes were created with the results of the Vitek Bacillus Biochemical Card test, and random amplified polymorphic DNA (RAPD) patterns were generated by arbitrarily primed PCR. Two clones of the 15 B. cereus isolates recovered from patients were identified (clone A from 2 patients and clone B from 13 patients), and all 29 isolates of B. cereus recovered from 70 or 95% ethyl alcohol in the hospital or in the factory belonged to clone B. The antibiotype and RAPD pattern of the B. sphaericus isolate from the patient were different from those of isolates from the factory. Our data show that the pseudoepidemic was caused by a clone (clone B) of B. cereus from contaminated 70% ethyl alcohol used in the hospital, which we successfully traced to preexisting contaminated 95% ethyl alcohol from the supplier, and by another clone (clone A) without an identifiable source.

An outbreak of Bacillus cereus respiratory tract infections on a neonatal unit due to contaminated ventilator circuits.

An outbreak of Bacillus cereus respiratory tract infections affecting six ventilated preterm neonates over a two-week period is described. Reusable ventilator circuits were identified as the cause of the outbreak. Ordinarily these were reprocessed on the Neonatal Unit (NNU), first through a washing machine and then through a low-temperature steam (LTS) disinfector. The onset of the outbreak coincided with a breakdown of the LTS facility, which necessitated sending the washed circuits off site for LTS disinfection. The washing machine was shown to be contaminated with the same serovars of B. cereus as those isolated from patients. Two critical steps in the off site LTS disinfection process allowed exsporulation and multiplication of B. cereus: the circuits were inadequately dried after processing, whilst return of the moist circuits to the NNU was often delayed. The outbreak was terminated by withdrawal of the heat-disinfected ventilator circuits. This outbreak emphasizes the need for high standards where medical equipment is reprocessed, especially for use in vulnerable patients.

Dissemination of Bacillus cereus in an intensive care unit.

OBJECTIVE: To report the contamination of ventilator equipment with Bacillus cereus and to outline the measures taken to trace the source of the organism. DESIGN: A prospective survey of all intensive care unit patients who were culture-positive for B cereus and obtaining of environmental cultures of the cleaning and assembly area of the respiratory services division between October 1991 and September 1992. SETTING AND PATIENTS: Ventilated patients from a 16-bed medical and surgical intensive care unit (ICU) in a 1,000-bed adult tertiary care hospital. INTERVENTIONS AND RESULTS: From October 1991 to April 1992, B cereus colonized the ventilator circuitry of patients in the ICU. One of two washer/decontaminators in the cleaning and assembly area of the respiratory services division was found to yield the microorganism consistently from the water intake port. The design of the machine precluded easy decontamination of the port with 2% glutaraldehyde and a second outbreak occurred. Following the second outbreak, aqueous chlorhexidine in a final concentration of 0.05% was added to the first of two pasteurization cycles in an attempt to achieve sporicidal activity. This ended the outbreak. Sixty-two patients became colonized with the organism including two with nonfatal Bacillus sepsis and one death due to pneumonia associated with the organism. CONCLUSION: This experience emphasizes the importance of obtaining cultures of machine parts to identify the source of contamination and thereby direct control measures. Use of chlorhexidine gluconate at high temperatures effectively eradicated B cereus from ventilator circuitry in a practical and cost-effective manner.

Bacillus cereus meningitis in two neurosurgical patients: an investigation into the source of the organism.

Two patients developed Bacillus cereus meningitis following neurosurgery. During the subsequent investigation into the source of the organism, linen was discovered to be heavily contaminated with B. cereus. No other prolific source of the organism was found. It seems probable that lint from contaminated fabric was the vehicle of transmission of the organism during extended surgery. Linen should be considered as a possible source of B. cereus infection.

Subclinical infection and transmission of Tyzzer's disease in rats.

Two isolates of Bacillus piliformis originally obtained from rats from Japan and Indiana were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Protein and antigen profiles revealed heterogeneity between the two isolates, demonstrating that more than one isolate of B. piliformis is capable of infecting rats. Results of parallel infection and transmission studies with the two isolates were almost identical. Orally inoculated rats remained asymptomatic; however, enzyme-linked immunosorbent assay results revealed a significant increase in serum antibodies to B. piliformis. Formalin-killed B. piliformis elicited no serum antibody response among rats inoculated orally, indicating that viable organisms, capable of replicating within the host, are needed to induce a systemic humoral response. Naive rats and weanling gerbils were housed on soiled bedding from the experimentally infected, asymptomatic, seropositive rats. Although gerbils showed no clinical signs or histopathologic evidence of Tyzzer's disease, rats housed on bedding collected 1 or 2 weeks postinoculation seroconverted and remained seropositive but asymptomatic throughout the study. These results demonstrate that subclinically infected rats are capable of transmitting B. piliformis to naive rats and suggest that the histopathologic evaluation of sentinel gerbils may not be an effective method for detecting all strains of B. piliformis.